Genome Editing in Animals Methods and Protocols

This volume details protocols that can be used for generation of knockout animals. Chapters guide the reader through basic protocols for three genome editing technologies, target design tools, and specific protocols for each animal. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Genome Editing in Animals: Methods and Protocols aims to ensure successful results in the further study of this vital field.

1.                  Construction and Evaluation of Zinc Finger Nucleases

Hiroshi Ochiai and Takashi Yamamoto

2.                  Current Overview of TALEN Construction Systems

Tetsushi Sakuma and Takashi Yamamoto

3.                  CRISPR/Cas9

Izuho Hatada and Takuro Horii

4.                  Computational Prediction of CRISPR/Cas9 Target Sites Reveals Potential Off-Target Risks in Human and Mouse

Qingbo Wang and Kumiko Ui-Tei

5.                  Genome Editing of Mouse by Cytoplasmic Injection

Takuro Horii and Izuho Hatada

6.                  Genome Editing in Mouse Zygotes and Embryonic Stem Cells by Introducing sgRNA/Cas9 Expressing Plasmids

7.                  Genome Editing in Mouse and Rat by Electroporation

Takehito Kaneko

8.                  Generation of Knock-in Mouse by Genome Editing

Wataru Fujii

9.                  Genome Editing of Rat

Takehito Kaneko

10.              Gene Targeting in Rabbits: Single-step Generation of Knockout Rabbits by Microinjection of CRISPR/Cas9 Plasmids

Yoshihiro Kawano and Arata Honda

11.              Genome Editing of Pig

Masahito Watanabe and Hiroshi Nagashima

12.              Genome Editing of Monkey

Zhen Liu, Yijun Cai, and Qiang Sun

13.              Genome Editing Mediated by Primordial Germ Cell in Chicken

Jae Yong Han and Hong Jo Lee

14.              CRISPR/Cas9-mediated Targeted Knock-in of Exogenous Reporter Genes in Zebrafish

Atsuo Kawahara

15.              Genome Editing of Medaka

Satoshi Ansai and Masato Kinoshita

16.              A Simple Protocol for Loss-of-Function Analysis in Xenopus tropicalis Founders using the CRISPR-Cas System

Yuto Sakane, Ken-ich T Suzuki, and Takashi Yamamoto

17.              Genome Editing of Silkworms

Takuya Tsubota and Hideki Sezutsu

18.              Genome Editing in the Cricket, Gryllus bimaculatus

Takahito Watanabe, Sumihare Noji, and Taro Mito

19.              Genome Editing of the Ascidian Ciona intestinalis with TALE Nuclease

Yasunori Sasakura, Keita Yoshida, and Nicholas Treen

20.              Genome editing of C. elegans

Takuma Sugi